Single-Cell Data and Weighted Correlation Network Analysis Revealed the Regulatory Mechanisms of Macrophages in Carotid Plaques.

IF 3.6 3区 医学 Q2 IMMUNOLOGY
Journal of Immunology Research Pub Date : 2025-07-21 eCollection Date: 2025-01-01 DOI:10.1155/jimr/9987367
Yakun Ding, Xiaoyang Niu, Peng Guo, Bing Wang
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引用次数: 0

Abstract

Background: Macrophages play a critical role in carotid plaque. Understanding the mechanisms of carotid plaque formation based on macrophage heterogeneity could provide valuable insights for clinical intervention. Methods: Single-cell transcriptome and bulk RNA-seq data of carotid plaque were obtained from public databases. Weighted gene correlation network analysis (WGCNA) identified gene modules linked to unstable plaques. Macrophage marker genes were intersected with module genes of WGCNA, followed by using randomForest and LASSO regression to pinpoint key genes. Quantitative real-time PCR (qRT-PCR) and Western blot were used to verify the regulation of key genes at the cellular level. The correlation between the key genes and inflammatory phenotypes was examined by single-sample gene set enrichment analysis (ssGSEA). Results: Single-cell clustering revealed major cellular subpopulations, with elevated macrophage infiltration in carotid plaque. Six key macrophage-associated genes (ADPGK, ATP6V1F, CX3CR1, MYO9B, RNF135, and SLC7A8) were discovered. The qRT-PCR results demonstrated upregulation of ADPGK, ATP6V1F, and RNF135 genes in vascular smooth muscle cells (VSMCs) treated with oxidized low-density lipoprotein (ox-LDL), except for CX3CR1, which was downregulated. Protein expression results showed that expressions of ADPGK, ATP6V1F, RNF135, and SLC7A8 were significantly elevated in the ox-LDL-VSMC group. In addition, most of the immune cells showed significant differences between the unstable arterial plaque group and the control group. Conclusion: This study discovered potential biomarkers that affected carotid plaque progression and macrophage regulation at the single-cell level, and examined their regulatory roles in immune regulation, programed cell death (PCD), and inflammatory factor modulation.

单细胞数据和加权相关网络分析揭示了巨噬细胞在颈动脉斑块中的调节机制。
背景:巨噬细胞在颈动脉斑块中起关键作用。了解基于巨噬细胞异质性的颈动脉斑块形成机制可以为临床干预提供有价值的见解。方法:从公共数据库获取颈动脉斑块单细胞转录组和大量RNA-seq数据。加权基因相关网络分析(WGCNA)确定了与不稳定斑块相关的基因模块。将巨噬细胞标记基因与WGCNA模块基因交叉,利用randomForest和LASSO回归找出关键基因。采用实时荧光定量PCR (qRT-PCR)和Western blot技术验证关键基因在细胞水平上的调控作用。通过单样本基因集富集分析(ssGSEA)检测关键基因与炎症表型之间的相关性。结果:单细胞聚集显示主要的细胞亚群,颈动脉斑块中巨噬细胞浸润升高。发现6个关键巨噬细胞相关基因(ADPGK、ATP6V1F、CX3CR1、MYO9B、RNF135和SLC7A8)。qRT-PCR结果显示,在氧化低密度脂蛋白(ox-LDL)处理的血管平滑肌细胞(VSMCs)中,除CX3CR1下调外,ADPGK、ATP6V1F和RNF135基因均上调。蛋白表达结果显示,ox-LDL-VSMC组ADPGK、ATP6V1F、RNF135、SLC7A8的表达显著升高。此外,大多数免疫细胞在不稳定动脉斑块组与对照组之间表现出显著差异。结论:本研究发现了在单细胞水平上影响颈动脉斑块进展和巨噬细胞调节的潜在生物标志物,并研究了它们在免疫调节、程序性细胞死亡(PCD)和炎症因子调节中的调节作用。
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来源期刊
CiteScore
6.90
自引率
2.40%
发文量
423
审稿时长
15 weeks
期刊介绍: Journal of Immunology Research is a peer-reviewed, Open Access journal that provides a platform for scientists and clinicians working in different areas of immunology and therapy. The journal publishes research articles, review articles, as well as clinical studies related to classical immunology, molecular immunology, clinical immunology, cancer immunology, transplantation immunology, immune pathology, immunodeficiency, autoimmune diseases, immune disorders, and immunotherapy.
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